scholarly journals Mapping Protein Structure Changes with Cysteine Labeling Kinetics Measured by Mass Spectrometry

2013 ◽  
Vol 104 (2) ◽  
pp. 168a
Author(s):  
Brian C. Chase ◽  
Joe Swift ◽  
J. David Pajerowski ◽  
Hsin-Yao Tang ◽  
David Speicher ◽  
...  
2012 ◽  
Vol 102 (3) ◽  
pp. 608a
Author(s):  
Brian C. Chase ◽  
J. David Pajerowski ◽  
Diego Pantano ◽  
Hsin-Yao Tang ◽  
David Speicher ◽  
...  

2019 ◽  
Vol 26 (1) ◽  
pp. 35-43 ◽  
Author(s):  
Natalie K. Garcia ◽  
Galahad Deperalta ◽  
Aaron T. Wecksler

Background: Biotherapeutics, particularly monoclonal antibodies (mAbs), are a maturing class of drugs capable of treating a wide range of diseases. Therapeutic function and solutionstability are linked to the proper three-dimensional organization of the primary sequence into Higher Order Structure (HOS) as well as the timescales of protein motions (dynamics). Methods that directly monitor protein HOS and dynamics are important for mapping therapeutically relevant protein-protein interactions and assessing properly folded structures. Irreversible covalent protein footprinting Mass Spectrometry (MS) tools, such as site-specific amino acid labeling and hydroxyl radical footprinting are analytical techniques capable of monitoring the side chain solvent accessibility influenced by tertiary and quaternary structure. Here we discuss the methodology, examples of biotherapeutic applications, and the future directions of irreversible covalent protein footprinting MS in biotherapeutic research and development. Conclusion: Bottom-up mass spectrometry using irreversible labeling techniques provide valuable information for characterizing solution-phase protein structure. Examples range from epitope mapping and protein-ligand interactions, to probing challenging structures of membrane proteins. By paring these techniques with hydrogen-deuterium exchange, spectroscopic analysis, or static-phase structural data such as crystallography or electron microscopy, a comprehensive understanding of protein structure can be obtained.


1994 ◽  
pp. 199-242 ◽  
Author(s):  
Amina Woods ◽  
Rong Wang ◽  
Marc Chevrier ◽  
Tim Cornish ◽  
Cathy Wolkow ◽  
...  

2019 ◽  
Vol 2019 ◽  
pp. 1-7 ◽  
Author(s):  
Longchun Li ◽  
Fan Wu ◽  
Yu Chen ◽  
Lei Xu ◽  
Xiaochang Hao ◽  
...  

Reactive oxygen species play important effects on organisms not only in vivo but also in vitro. The atomic oxygen radical anion (O-) has shown extremely high oxidation and reactivity towards small molecules of hydrocarbons. However, the O- effects on cells of microorganisms are scarcely investigated. This work showed the evidence that O- could react quickly with microorganisms (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Aspergillus niger, Saccharomyces cerevisiae, and Actinomycetes (5046)) and damaged the cell walls seriously as well as their intrinsic structures, arising a fast and irreversible inactivation. SEM and TEM micrographs were used to reveal the structure changes of cells before and after reacting with O- radicals. The inactivation efficiencies of the microorganisms depended on the O- intensity, the initial population of microorganisms, the exposed area, the environment, and the microorganisms’ types. Over 99% reduction of an initial 1.0×107 colony-forming unit (cfu), E. coli population only required less than 2 minutes while exposed to a 0.23 μA/cm2 O- flux under dry argon atmosphere (30°C, 1 atm). The observation of anionic intermediates (CO-, CO2-, H2O-, and anionic hydrocarbons) by time-of-flight (TOF) mass spectrometry and the neutral volatile products (CO, CO2, and H2O) by quadrupole mass spectrometry (Q-MS) provided an evidence of the reactions of O- with hydrocarbon bonds of the microorganisms. The inactivation mechanism of microorganisms induced by O- was discussed.


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